Programa de Pós-Graduação em Ciências Biológicas: Fisiologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil.
Programa de Pós-Graduação em Farmácia, Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil.
J Oral Pathol Med. 2023 Oct;52(9):877-884. doi: 10.1111/jop.13475. Epub 2023 Aug 7.
Considering that microRNAs (miRNAs), extracellular vesicles and particles (EVPs) and the amyloid precursor protein (APP) processing have been shown to be altered in oral squamous cells carcinoma (OSCC), it is possible that miRNAs that target APP processing pathways in EVPs are impacted in tumor cells. Our aim was to evaluate miRNAs that target APP itself or disintegrin and metalloproteinase domain 10 (ADAM10), which generate a trophic compound, sAPPα, in EVPs derived from OSCC cell lines, an aggressive and non-invasive, compared to normal keratinocytes.
We used two OSCC cell lines, an aggressive human oral squamous cell carcinoma cell line (SCC09) and a less aggressive cell line (CAL27) compared with a keratinocyte lineage (HaCaT). Cells were maintained in cell media, from which we isolated EVPs. EVPs were evaluated regarding their size and concentration using Nanotracking Analysis. We measured the levels of miRNAs which had as potential downstream target APP or ADAM10, specifically miR-20a-5p, miR-103a-3p, miR-424-5p, miR-92b-3p, miR-31-5p, and miR-93-5.
There were no differences on size distributions and concentration of isolated EVPs. OSCC cell lines-derived EVPs miR-20a-5p, miR-92b-3p, and miR-93-5p were upregulated in comparison to HaCaT-derived EVPs; while miR-31-5p was reduced in EVPs obtained from CAL27 cells.
Our results indicate changes in miRNAs that target APP machinery processing in EVPs derived from OSCC cell lines of different aggressiveness, which may be involved with abnormal miRNA expression in OSCC tissue and/or releasing tumor suppressor miRNA.
考虑到 microRNAs（miRNAs）、细胞外囊泡和颗粒（EVP）以及淀粉样前体蛋白（APP）的处理在口腔鳞状细胞癌（OSCC）中已经发生改变，因此靶向 EVP 中 APP 处理途径的 miRNAs 可能在肿瘤细胞中受到影响。我们的目的是评估靶向 APP 本身或解整合素金属蛋白酶域 10（ADAM10）的 miRNAs，后者在源自 OSCC 细胞系的 EVP 中产生一种营养化合物 sAPPα，与正常角质形成细胞相比，这些细胞系具有侵袭性和非侵袭性。
我们使用了两种 OSCC 细胞系，一种是侵袭性人口腔鳞状细胞癌细胞系（SCC09），另一种是侵袭性较弱的细胞系（CAL27），与角质形成细胞系（HaCaT）相比。细胞在细胞培养基中培养，从培养基中分离 EVP。使用纳米跟踪分析评估 EVP 的大小和浓度。我们测量了具有 APP 或 ADAM10 下游靶标的 miRNAs 的水平，特别是 miR-20a-5p、miR-103a-3p、miR-424-5p、miR-92b-3p、miR-31-5p 和 miR-93-5p。
分离的 EVP 的大小分布和浓度没有差异。与 HaCaT 衍生的 EVP 相比，OSCC 细胞系衍生的 EVPs 中的 miR-20a-5p、miR-92b-3p 和 miR-93-5p 上调；而源自 CAL27 细胞的 EVPs 中 miR-31-5p 减少。
我们的结果表明，不同侵袭性 OSCC 细胞系衍生的 EVP 中靶向 APP 机制处理的 miRNAs 发生变化，这可能与 OSCC 组织中异常 miRNA 表达和/或释放肿瘤抑制 miRNA 有关。
